Novel Approach to Designing Primers for Identification and Distinction of the Human Pathogenic Fungi
            <i>Coccidioides immitis</i>
            and
            <i>Coccidioides posadasii</i>
            by PCR Amplification

Umeyama, Takashi; Sano, Ayako; Kamei, Katsuhiko; Niimi, Masakazu; Nishimura, Kazuko; Uehara, Yoshimasa

doi:10.1128/jcm.44.5.1859-1862.2006

Cited by 41 publications

(40 citation statements)

References 11 publications

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“…All strains belonged to the fungal collection of the Specialized Medical Mycology Center (CEMM, Federal University of Ceará, Brazil). The procedures for identification of the fungi included classic mycological analysis, as described by Brilhante et al (19), and a PCR assay (20). All procedures were performed in a class II biological safety cabinet in a biosafety level 3 laboratory.…”

Section: Methodsmentioning

confidence: 99%

Effect of Farnesol on Growth, Ergosterol Biosynthesis, and Cell Permeability in Coccidioides posadasii

Brilhante

Lima

Caetano

et al. 2013

Antimicrob Agents Chemother

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Coccidioidomycosis is a systemic mycosis caused by the dimorphic fungi Coccidioides spp. The treatment for chronic and/or disseminated coccidioidomycosis can be prolonged and complicated. Therefore, the search for new drugs is necessary. Farnesol is a precursor in the sterol biosynthesis pathway that has been shown to present antifungal activity. Thus, the objective of this study was to evaluate the in vitro antifungal activity of farnesol alone and in combination with antifungal agents against clinical and environmental strains of Coccidioides posadasii as well as to determine their effect on the synthesis of ergosterol and on cell permeability. This study employed the broth macrodilution method to determine the MIC of farnesol against 18 strains of C. posadasii. Quantification of ergosterol was performed with 10 strains of C. posadasii after exposure to subinhibitory concentrations of farnesol. Finally, the activity of farnesol was evaluated in the presence of osmotic stress, induced by the addition of NaCl to the culture medium, during the susceptibility tests. The results showed that farnesol exhibited low MICs (ranging from 0.00171 to 0.01369 mg/liter) against all tested strains. The combination of farnesol with the antifungals showed synergistic effects (fractional inhibitory concentration index [FICI] < 0.5). As for the ergosterol quantification, it was observed that exposure to subinhibitory concentrations of farnesol decreased the amount of ergosterol extracted from the fungal cells. Furthermore, farnesol also showed lower MIC values when the strains were subjected to osmotic stress, indicating the action of this compound on the fungal membrane. Thus, due to the high in vitro antifungal activity, this work brings perspectives for the performance of in vivo studies to further elucidate the effects of farnesol on the host cells.

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Section: Methodsmentioning

confidence: 99%

Effect of Farnesol on Growth, Ergosterol Biosynthesis, and Cell Permeability in Coccidioides posadasii

Brilhante

Lima

Caetano

et al. 2013

Antimicrob Agents Chemother

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“…Se resembraron los cultivos para extraer su ADN y se amplificó una región con los oligonucleótidos Coi9-1F y Coi9-1R diseñados por Umeyama 14 .…”

Section: Discussionunclassified

Coccidioidomicosis cutánea: Comunicación de seis casos mexicanos

Moreno‐Coutiño¹,

Arce-Ramírez²,

Medina³

et al. 2015

Rev. chil. infectol.

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“…Polymerase chain reaction (PCR) assays targeting conserved or variable regions of the ribosomal DNA to detect and identify Coccidioides spp. have been described and found to be sensitive and specific 2,12,15,16,18,21,27,30,32 . Molecular phylogenetic methods based on single-nucleotide polymorphisms and microsatellite repeats have established a new species, Coccidioides posadasii, which is morphologically indistinguishable from C. immitis 12,19,30 .…”

Section: Introductionmentioning

confidence: 99%

Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil

Cavalcanti

Vidal

Sousa

et al. 2013

Rev. Inst. Med. trop. S. Paulo

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SUMMARYCoccidioidomycosis is an emerging fungal disease in Brazil; adequate maintenance and authentication of Coccidioides isolates are essential for research into genetic diversity of the environmental organisms, as well as for understanding the human disease. Seventeen Coccidioides isolates maintained under mineral oil since 1975 in the Instituto de Medicina Tropical de São Paulo (IMTSP) culture collection, Brazil, were evaluated with respect to their viability, morphological characteristics and genetic features in order to authenticate these fungal cultures. Only five isolates were viable after almost 30 years, showing typical morphological characteristics, and sequencing analysis using Coi-F and Coi-R primers revealed 99% identity with Coccidioides genera. These five isolates were then preserved in liquid nitrogen and sterile water, and remained viable after two years of storage under these conditions, maintaining the same features.

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Novel Approach to Designing Primers for Identification and Distinction of the Human Pathogenic Fungi Coccidioides immitis and Coccidioides posadasii by PCR Amplification

Cited by 41 publications

References 11 publications

Effect of Farnesol on Growth, Ergosterol Biosynthesis, and Cell Permeability in Coccidioides posadasii

Effect of Farnesol on Growth, Ergosterol Biosynthesis, and Cell Permeability in Coccidioides posadasii

Coccidioidomicosis cutánea: Comunicación de seis casos mexicanos

Viability and molecular authentication of Coccidioides spp. isolates from the Instituto de Medicina Tropical de São Paulo culture collection, Brazil

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